Multicenter Comparison Trial of DNA Extraction Methods and PCR Assays for Detection of Chlamydia pneumoniae in Endarterectomy Specimens

doi:10.1128/JCM.39.2.519-524.2001

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Journal of Clinical Microbiology, February 2001, p. 519-524, Vol. 39, No. 2
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.2.519-524.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Multicenter Comparison Trial of DNA Extraction Methods and PCR Assays for Detection of Chlamydia pneumoniae in Endarterectomy Specimens

Petra Apfalter,¹^,^* Francesco Blasi,² Jens Boman, Charlotte A. Gaydos,⁴ Michael Kundi,⁵ Matthias Maass,⁶ Athanasios Makristathis,¹ Adam Meijer,⁷ Reinhard Nadrchal,¹ Kenneth Persson,⁸ Manfred L. Rotter,¹ C. Y. William Tong,⁹^, Gerold Stanek,¹⁰ and Alexander M. Hirschl¹

Department of Clinical Microbiology,¹ and Department of Infection Immunology,¹⁰ Hygiene-Institute, and Department of Occupational and Social Hygiene, Institute of Environmental Health,⁵ University of Vienna, Vienna, Austria; Institute of Respiratory Diseases, IRCCS Policlinico, University of Milan, Milan, Italy²; Department of Clinical Virology, Umeå University Hospital, Umeå, Sweden³; Division of Infectious Diseases, Department of Medicine, The Johns Hopkins University, Baltimore, Maryland⁴; Institute of Medical Microbiology, Medical University of Lübeck, Lübeck, Germany⁶; Research Laboratory for Infectious Diseases, National Institute of Public Health and the Environment, Bilthoven, The Netherlands⁷; Department of Clinical Virology, Malmö General Hospital, University of Lund, Lund, Sweden⁸; and Department of Medical Microbiology and Genitourinary Medicine, University of Liverpool, Liverpool, United Kingdom⁹

Received 16 August 2000/Returned for modification 11 November 2000/Accepted 25 November 2000

The reported rate of detection of Chlamydia pneumoniae DNA within atherosclerotic lesions by PCR varies between 0 and 100%.In this study, identical sets of coded experimental atheroma samples(n = 15) and spiked controls (n = 5) were analyzed by 16 testmethods in nine centers by means of PCR. The positive controlswere correctly identified to levels of 1, 0.1, and 0.01 inclusionbodies of C. pneumoniae/ml of tissue homogenate by 16 (100%),11 (69%), and 3 (19%) of the test methods, respectively. Threeout of 16 negative controls (19%) were rated positive. Positivityrates for atheroma samples varied between 0 and 60% for the differenttest methods, with the maximum concordant result for positivitybeing only 25% for one carotid artery sample. There was no consistentpattern of positive results among the various laboratories, andthere was no correlation between the detection rates and the sensitivityof the assayused.

^* Corresponding author. Mailing address: Department of Clinical Microbiology, Hygiene-Institute, University of Vienna, ViennaUniversity Hospital, Währinger Gürtel 18-20/5P, A-1090 Vienna,Austria. Phone: 43 (1) 40400-5151. Fax: 43 (1) 40400-5228. E-mail:Petra.Apfalter{at}akh-wien.ac.at.

Present address: Department of Infection, Virology Section, St. Thomas' Hospital, London SE 1 7EH, UnitedKingdom.

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